The Growth and Maintenance of Tissue–cell Cultures in Free Gas Exchange With the Atmosphere1

Leibovitz, Albert

doi:10.1093/oxfordjournals.aje.a120336

Cited by 236 publications

(131 citation statements)

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“…38,39 These observations led to the development of a culture medium not requiring a complex buffer system. [40][41][42] So far, no experience has been obtained with (human) hematopoietic cells. Table 4 shows that indeed a better control of pH was found using L15 medium, although the range was still very wide.…”

Section: Discussionmentioning

confidence: 99%

Storage of unprocessed G-CSF-mobilized whole blood in a modified Leibovitz's L15 medium preserves clonogenic capacity for at least 7 days

Kreuk

Jonkhoff

Zevenbergen

et al. 2001

Bone Marrow Transplant

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Summary:Autologous stem cell transplantation using unprocessed, G-CSF-mobilized whole blood (WB) is a simple, cost-reducing procedure and supports high-dose chemotherapy regimens not exceeding 72 h. Thereafter, clonogenic capacity rapidly decreases if routine anticoagulants are used for storage. In order to increase clinical applicability, we investigated the requirements for optimal preservation of unprocessed WB for 7 days. During storage at 22؇C in CPDA-1, a decrease in pH was noted, which was at least partially responsible for the low recovery of clonogenic cells. Subsequently, WB cells were stored in various cell culture media (RPMI 1640, ␣-MEM, X-VIVO15, CellGro SCGM and Leibovitz's L15 medium) containing either serum, serum-free substitutes or no additives. Leibovitz's L15 showed significantly better CFU-GM recoveries than the other media. Using a calcium-free modification of L15 medium (added 3:10 to WB), 94 ؎ 24% of CD34 ؉ cells, 41 ؎ 14% of BFU-E, 56 ؎ 17% CFU-GM and 90 ؎ 14% of LTC-IC were preserved during storage for 7 days at 22؇C. Storage at 4؇C was also feasible, but showed less optimal recoveries of 52 ؎ 29% (CD34), 32 ؎ 10% (BFU-E), 13 ؎ 7% (CFU-GM) and 58 ؎ 9% (LTC-IC). The expression of CD38, Thy-1, c-kit, AC133, L-selectin and CXCR4 on CD34-positive cells remained unchanged. In conclusion, a modified Leibovitz's L15 medium better meets the metabolic requirements of a high-density cell culture and allows safe storage of G-CSF mobilized WB for at least 7 days. The results encourage further exploration of WB transplants stored for 7 days for clinical use. Bone Marrow Transplantation (2001) 28, 145-155.

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Section: Discussionmentioning

confidence: 99%

Storage of unprocessed G-CSF-mobilized whole blood in a modified Leibovitz's L15 medium preserves clonogenic capacity for at least 7 days

Kreuk

Jonkhoff

Zevenbergen

et al. 2001

Bone Marrow Transplant

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“…The cell suspension was diluted to 250 000 cells/ml with L~15 medium (Leibovitz 1963) (Life Technologies, Paisley, Strathclyde, UK) containing 26 niM sodium bicarbonate, and 200 jal aliquots were placed on glass coverslips (24 x 24 mm) in six-well tissue culture plates (Costar, Cambridge, MA, USA). Cells were cultured for 90 min at 25 °C, 5% C 0 2 to allow attachment of the cells to the coverslip, after which 2-8 ml L-15 medium containing 26 mM sodium bicarbonate and horse serum (Gibco, Paisley, Strathclyde, UK; final concen tration 5% (v/v)) was added.…”

Section: Pituitary Cell Dispersion and G Nrh Receptor Localisationmentioning

confidence: 99%

Gonadotrophs but not somatotrophs carry gonadotrophin-releasing hormone receptors: receptor localisation, intracellular calcium, and gonadotrophin and GH release

Bosma¹,

Kolk²,

Rebers³

et al. 1997

Journal of Endocrinology

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Gonadotrophs are the primary target cells for GnRH in the pituitary. However, during a limited period of neonatal life in the rat, lactotrophs and somatotrophs respond to GnRH as well. Also, in the adults of a number of teleost fishes (e.g. carp, goldfish, and tilapia but not trout), GnRH is a potent GH secretagogue. In studying hypophysiotrophic actions of the two forms of GnRH present in the African catfish (Clarias gariepinus), chicken GnRH-II ([His5,Trp7,TyrK ] GnRH; cGnRH-II) and catfish GnRH ([His5,Asn8]GnRH; cfGnRH), we have investigated the effects of G nRH on catfish gonadotrophs and somato trophs. G nRH binding was examined by incubating dispersed pituitary cells attached to coverslips with 125I~ labelled [D-Argr\T rp 7,LeuH ,Pro9-Net]GnRH (sGnRHa), a salmon G nRH analogue with high affinity for the G nRH receptor. Following fixation and immunohistochemistry using antisera against catfish LH and GH, 125I-labelled sGnRHa was localised autoradiographically and silver grains were quantified on gonadotrophs and somatotrophs. Specific binding of ' I-labelled sGnRHa was restricted to gonadotrophs. Both cfGnRH andcGnRH~II dose-dependently inhibited I-labelled sGnRHa binding to gonadotrophs. To substantiate the localisation of functional G nR H receptors, the effects of cfGnRH and cG nR H -II on the cytosolic free calcium concentration ([Ca ' were examined in Fura-2-loaded somatotrophs and gonadotrophs. GnRH-induced increases in [Ca2+]i appeared to be confined to gonadotrophs, in which both endogenous GnRHs caused a single and transient increase in [Ca2*]}. The amplitude of this [Ca2+]j transient depended on the G nR H dose and correlated well with the G nR H s1 effect on LH release. In vivo experiments demonstrated that G nR H treatments which markedly elevated plasma LH levels had no effect on plasma GH levels, while a dopamine agonist (apomorphine) significantly elevated plasma GH levels. W e con clude that the two endogenous forms of G nR H in the African catfish are not directly involved in the regulation of the release of GH, suggesting that GnRHs cannot be considered as GH secretagogues in teleosts in general.

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“…Vero cells were used with medium (Leibovitz, 1963) containing 5 % inactivated fetal calf serum (FCS) and 50,g/ml gentamicin. Virus stocks consisted of the supernatant fluids from Vero cell cultures infected with RVF 1678/78 stored in small volumes at -70 'C.…”

Section: Cytopathic Effect Neutralization (Cpent) Testsmentioning

confidence: 99%

Comparison of techniques for demonstrating antibodies to Rift Valley fever virus

Swanepoel

Struthers

Erasmus

et al. 1986

J. Hyg.

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SUMMARYNine serological techniques were compared by monitoring the response to infection with Rift Valley fever (RVF) virus in three sheep. Antibodies were monitored daily for the first 14 days after infection, then weekly and later fortnightly up to week 24. The earliest antibody response was detected in one sheep on day 3 by a plaque reduction neutralization test, and by day 6 antibodies were demonstrable in all three sheep by haemagglutination-inhibition, reversed passive haemagglutination-inhibition, immunodiffusion, indirect immunofluorescence (IF), enzyme-linked immunosorbent assay and neutralization ofcytopathic effect in cell cultures. Antibodies were demonstrable by complement fixation on day 8 at the earliest. IF and the two neutralization techniques produced the highest titres, but all tests could be used satisfactorily for the serological diagnosis of RVF. Inactivated antigen could be used for all except the neutralization tests. A radioimmunoassay technique using l251-labelled staphylococcal protein A detected antibodies on day 8 at the earliest and produced lower mean titres than some of the other techniques. This was probably because sheep immunoglobulins bind protein A poorly.

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The Growth and Maintenance of Tissue–cell Cultures in Free Gas Exchange With the Atmosphere1

Cited by 236 publications

References 0 publications

Storage of unprocessed G-CSF-mobilized whole blood in a modified Leibovitz's L15 medium preserves clonogenic capacity for at least 7 days

Storage of unprocessed G-CSF-mobilized whole blood in a modified Leibovitz's L15 medium preserves clonogenic capacity for at least 7 days

Gonadotrophs but not somatotrophs carry gonadotrophin-releasing hormone receptors: receptor localisation, intracellular calcium, and gonadotrophin and GH release

Comparison of techniques for demonstrating antibodies to Rift Valley fever virus

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